Case report: Urolithiasis, nephrolithiasis and a urinary bladder malformation in a seven-month-old alpaca cria

Urolithiasis is a common condition in male small ruminants where predisposing factors have been identified. Occasionally, urolithiasis is diagnosed in South American camelids (SACs). However, nephrolithiasis is rarely diagnosed in ruminants. To our knowledge, this is the first report focusing on a combined appearance of nephrolithiasis and urolithiasis in an alpaca cria. A 7-month-old alpaca cria suffering from impaired urinary flow was presented for examination. On admission, the alpaca had a wet prepuce and showed a standing posture with a wide-based stance. Ultrasonographic examination of the abdomen showed a distended bladder. Clinical chemistry revealed azotemia and hypophosphatemia. After the first examination, repeated urination was observed. Conservative therapy using antibiotics, anti-inflammatory and spasmolytic drugs was started with the suspected diagnosis of urinary calculus. During the first 24 h, plasma concentrations of creatinine and urea decreased, but increased again during the following days. During the second day after admission, urination was not observed for 16 h while the concentration of urea and creatinine further increased. Therefore, the animal was euthanized due to financial concerns of the owner. Necropsy revealed that calculi were located in the left kidney as well as in the urethra. In addition, the animal exhibited uroperitoneum. The urinary bladder was intact, moderately distended with urine and showed a malformation, which was covered with a translucent mucosal membrane. Histologic examination revealed that this malformation was a bladder diverticulum. The extent to which the unilateral nephroliths affected the general condition and renal function of the animal is unclear, since the uroliths also cause azotemia, and abdominal pain. Further studies are needed for a better understanding of obstructive urinary disease in SACs

Case report: Urolithiasis, nephrolithiasis and a urinary bladder malformation in a seven-month-old alpaca cria

Urolithiasis is a common condition in male small ruminants where predisposing factors have been identified. Occasionally, urolithiasis is diagnosed in South American camelids (SACs). However, nephrolithiasis is rarely diagnosed in ruminants. To our knowledge, this is the first report focusing on a combined appearance of nephrolithiasis and urolithiasis in an alpaca cria. A 7-month-old alpaca cria suffering from impaired urinary flow was presented for examination. On admission, the alpaca had a wet prepuce and showed a standing posture with a wide-based stance. Ultrasonographic examination of the abdomen showed a distended bladder. Clinical chemistry revealed azotemia and hypophosphatemia. After the first examination, repeated urination was observed. Conservative therapy using antibiotics, anti-inflammatory and spasmolytic drugs was started with the suspected diagnosis of urinary calculus. During the first 24 h, plasma concentrations of creatinine and urea decreased, but increased again during the following days. During the second day after admission, urination was not observed for 16 h while the concentration of urea and creatinine further increased. Therefore, the animal was euthanized due to financial concerns of the owner. Necropsy revealed that calculi were located in the left kidney as well as in the urethra. In addition, the animal exhibited uroperitoneum. The urinary bladder was intact, moderately distended with urine and showed a malformation, which was covered with a translucent mucosal membrane. Histologic examination revealed that this malformation was a bladder diverticulum. The extent to which the unilateral nephroliths affected the general condition and renal function of the animal is unclear, since the uroliths also cause azotemia, and abdominal pain. Further studies are needed for a better understanding of obstructive urinary disease in SACs

Expression levels of immune markers in Actinobacillus pleuropneumoniae infected pigs and their relation to breed and clinical symptoms

<p>Abstract</p> <p>Background</p> <p>In pigs little is known about the role of innate immune defence in bacterial infections of the respiratory tract, despite their major role in pig production. In the present study we characterized and compared <it>in vitro </it>and <it>in vivo </it>activation of immune markers of different pig breeds 7 days before, and 4 and 21 days after an experimental aerosol infection with <it>Actinobacillus (A.) pleuropneumoniae</it>.</p> <p>Results</p> <p><it>In vitro </it>stimulation of bronchoalveolar lavage fluid (BALF) and blood leukocytes with <it>A. pleuropneumoniae, Streptococcus suis</it>, PMA and LPS led to production of different amounts of H₂O₂, NO and TNF-α, depending on the stimulus, individual, breed and time of infection. Generally, significant responses to <it>in vitro </it>stimulation were observed only in blood leukocytes, whereas the alveolar macrophages showed a high basal activation. In addition, the production of haptoglobin and cytokines (TNF-α, IFN-γ and IL-10) <it>in vivo </it>was measured in plasma and BALF. Plasma haptoglobin levels mirrored the clinical manifestations at 4 days post-infection. In plasma and BALF TNF-α could not be detected, whereas variable levels of IFN-γ were found at pre- and post-infection times. IL-10 was found in some plasma but in none of the BALF samples. The different expression levels in individuals within the breeds correlated for some markers with the severity of clinical manifestations, e.g. H₂O_2,plasma haptoglobin and BALF IFN-γ for German Landrace pigs.</p> <p>Conclusion</p> <p>Our findings revealed differences in the activation of the immune markers with respect to infection time, individuals and breeds. Moreover, results showed different correlation grades between the immune markers produced <it>in vitro </it>or <it>in vivo </it>and the clinical manifestations. Further analyses will have to show whether these markers may serve as correlates of protection against porcine respiratory infections.</p

Evaluation of a Method for Standardized Antimicrobial Susceptibility Testing with Mycoplasma hyorhinis Field Isolates

Organizations like the Clinical and Laboratory Standards Institute (CLSI) or the European Committee of Antimicrobial Susceptibility Testing (EUCAST) provide standardized methodologies for antimicrobial susceptibility testing of a wide range of nonfastidious and fastidious bacteria, but so far not for Mycoplasma spp. of animal origin. Recently, a proposed method for the standardized broth microdilution testing of Mycoplasma hyorhinis using commercial Sensititre microtiter plates was presented. In this study, we evaluated this broth microdilution method with 37 field isolates and tested their susceptibility toward the following antimicrobial agents: doxycycline, enrofloxacin, erythromycin, florfenicol, gentamicin, marbofloxacin, tetracycline, tiamulin, tilmicosin, tulathromycin, and tylosin. The isolates originated from different countries, isolation sites, and years. The broth microdilution method was carried out using a modified Friis broth as the culture and test medium. For macrolides and lincosamides, a bimodal distribution with elevated MIC values could be observed for almost half of the tested field isolates, deducing reduced susceptibility toward these substances. With a recently published protocol, we were able to test a variety of field isolates, and consistent data could be obtained. Using this method, monitoring studies of Mycoplasma hyorhinis isolates can be carried out in a comparable manner, and the observed susceptibility profiles can be screened for possible changes in MIC values in the future

Experimental Actinobacillus pleuropneumoniae challenge in swine: comparison of computed tomographic and radiographic findings during disease

BACKGROUND: In pigs, diseases of the respiratory tract like pleuropneumonia due to Actinobacillus pleuropneumoniae (App) infection have led to high economic losses for decades. Further research on disease pathogenesis, pathogen-host-interactions and new prophylactic and therapeutic approaches are needed. In most studies, a large number of experimental animals are required to assess lung alterations at different stages of the disease. In order to reduce the required number of animals but nevertheless gather information on the nature and extent of lung alterations in living pigs, a computed tomographic scoring system for quantifying gross pathological findings was developed. In this study, five healthy pigs served as control animals while 24 pigs were infected with App, the causative agent of pleuropneumonia in pigs, in an established model for respiratory tract disease. RESULTS: Computed tomographic (CT) findings during the course of App challenge were verified by radiological imaging, clinical, serological, gross pathology and histological examinations. Findings from clinical examinations and both CT and radiological imaging, were recorded on day 7 and day 21 after challenge. Clinical signs after experimental App challenge were indicative of acute to chronic disease. Lung CT findings of infected pigs comprised ground-glass opacities and consolidation. On day 7 and 21 the clinical scores significantly correlated with the scores of both imaging techniques. At day 21, significant correlations were found between clinical scores, CT scores and lung lesion scores. In 19 out of 22 challenged pigs the determined disease grades (not affected, slightly affected, moderately affected, severely affected) from CT and gross pathological examination were in accordance. Disease classification by radiography and gross pathology agreed in 11 out of 24 pigs. CONCLUSIONS: High-resolution, high-contrast CT examination with no overlapping of organs is superior to radiography in the assessment of pneumonic lung lesions after App challenge. The new CT scoring system allows for quantification of gross pathological lung alterations in living pigs. However, computed tomographic findings are not informative of the etiology of respiratory disease

Evaluation of the predictive value of tonsil examination by bacteriological culture for detecting positive lung colonization status of nursery pigs exposed to Actinobacillus pleuropneumoniae by experimental aerosol infection

Abstract Background Actinobacillus (A.) pleuropneumoniae is the causative agent of porcine pleuropneumonia. For control of the disease the detection of sub-clinically infected pigs is of major importance to avoid transmitting of subclinical infections. One method recommended is the testing of tonsillar samples for the presence of A. pleuropneumoniae. This is routinely done by PCR techniques. However, based upon PCR susceptibility testing and monitoring of resistance development is impossible. Therefore, in this study the informative values of bacteriological culture of tonsilar samples for the colonisation status of pigs were tested. In total, 163 German Landrace nursery pigs were experimentally exposed to A. pleuropneumoniae serotype 7 by aerosol and the rate of isolation from lung tissue and tonsils and the corresponding degree of lung lesions were investigated. Results Overall a significant correlation (p < 0.001) between degree of clinical disease, degree of lung alterations and degree of A. pleuropneumoniae isolation from tonsillar and lung tissue after exposure was detected. Of these animals tested, 74.8% were tested positive in tonsillar and lung samples, 7.4% remained completely negative and in 4.3% the tonsils were tested positive despite negative isolation results from lung tissue. In 13.5% of the pigs A. pleuropneumoniae could be isolated in lung tissue but not in tonsillar samples. In 36.4% of these animals a heavy colonization of the lungs and in 40.9% moderate to severe lung alterations were proven. Hence, the diagnostic sensitivity for the detection of a positive colonization status of the pigs by bacterial culture examination of tonsillar samples was 84.7%, the diagnostic specificity was 66.7% and the predictive values were 94.6% (positive) and 35.3% (negative). The overall sensitivity for A. pleuropneumoniae exposure was 78.2% (tonsils) and 88.0% (lung tissue). Conclusions In conclusion, tonsil examination alone for the detection of a positive colonization status of pigs performed might lead to false negative results as lungs might be heavily colonized despite negative tonsillar isolation results. Therefore culture of tonsillar samples should not be the sole test for the confirmation of a pigs’ status but used in combination with methods also evaluating the colonization status of the lower respiratory tract

Sero- and apx-typing of German Actinobacillus pleuropneumoniae field isolates from 2010 to 2019 reveals a predominance of serovar 2 with regular apx-profile

Serotyping is the most common method to characterize field isolates of Actinobacillus (A.) pleuropneumoniae, the etiological agent of porcine pleuropneumonia. Based on serology, many farms seem to be infected and antibodies against a wide variety of serovars are detectable, but, so far it is unknown to what degree respective serovars contribute to outbreaks of clinical manifest disease. In this study, 213 German A. pleuropneumoniae field isolates retrieved for diagnostic purposes from outbreaks of porcine pleuropneumonia between 2010 and 2019 were genetically serotyped and analyzed regarding their apx-toxin gene profile using molecular methods. Serotyping revealed a prominent role of serovar 2 in clinical cases (64% of all isolates) and an increase in the detection of this serovar since 2010 in German isolates. Serovar 9/11 followed as the second most frequent serovar with about 15% of the isolates. Furthermore, very recently described serovars 16 (n = 2) and 18 (n = 8) were detected. Most isolates (93.4%) showed apx-profiles typical for the respective serovar. However, this does not hold true for isolates of serovar 18, as 75% (n = 6) of all isolates of this serovar deviated uniformly from the “typical” apx-gene profile of the reference strain 7311555. Notably, isolates from systemic lesions such as joints or meninges did not harbor the complete apxICABD operon which is considered typical for highly virulent strains. Furthermore, the extremely low occurrence (n = 1) of NAD independent (biovar II) isolates in German A. pleuropneumoniae was evident in our collection of clinical isolates

Efficacy of a one-shot marbofloxacin treatment on acute pleuropneumonia after experimental aerosol inoculation of nursery pigs

Abstract Background Porcine pleuropneumonia, caused by Actinobacillus pleuropneumoniae, is a bacterial respiratory disease of swine. Acute outbreaks of the disease are often accompanied by high mortality and economic losses. As severe cases of the disease frequently require parenteral antibiotic treatment of the animals, the efficacy of a single, high dose of marbofloxacin was compared to a three-time application of a dose of enrofloxacin under experimental conditions. Methods A blinded, controlled, randomized and blocked dose confirmation study was conducted to test the efficacy and safety of a single dose of 8 mg/kg marbofloxacin (160 mg/ml, Forcyl® Swine, Vetoquinol SA, France) to treat acute porcine pleuropneumonia after experimental aerosol inoculation of pigs with A. pleuropneumoniae serotype 2. The results were compared to a three consecutive day treatment of 2.5 mg/kg enrofloxacin and a mock (saline) treatment. Criteria for the assessment of efficacy were severity of lung lesions, bacteriological cure and the course of clinical disease after treatment. Results: Thirty six nursery pigs were divided into three treatment groups: marbofloxacin (T1), enrofloxacin (T2) and mock (T3). Statistically significant superiority (p < 0.05) of marbofloxacin and enrofloxacin compared to the mock-treated group was demonstrated for all efficacy criteria. The need of rescue euthanasia due to severity of symptoms was significantly reduced in both treatment groups (T1: 1 pig; T2: 0 pigs; vs. T3: 8 pigs). On day 6 after treatment initiation, clinical cure was observed in 10 (T1), 10 (T2) but only 1 of the piglets in T3. Extent of lung lesions (mean of lung lesion score T1: 3.9, T2: 6.0, T3: 21.1) and bacteriological isolation from lung tissue (on day 6 after treatment initiation: T1 = 0 pigs; T2 = 1 pig; T3 = all pigs) were also significantly reduced within both treatment groups. There were no adverse events linked to the drug administration and no injection site reactions were observed. Conclusions Both applied antimicrobial treatments were proven safe and efficacious for the treatment of acute porcine pleuropneumonia. No statistically significant differences were detected between the antibiotic treatments

Serotyping and pathotyping of Glaesserella parasuis isolated 2012–2019 in Germany comparing different PCR-based methods

Glaesserella parasuis is an important pathogen in swine production. It acts as a primary pathogen in systemic Glässer´s disease and as a secondary pathogen in Porcine Respiratory Disease Complex. In this study, a collection of 308 isolates from carrier animals and individuals with respiratory or Glässer´s disease isolated 2012–2019 in Germany was analysed. Isolates were characterized for serovar implementing two different PCR methods. Additionally, two different PCR methods for pathotyping isolates were applied to the collection and results compared. Serovar 6 (p &amp;lt; 0.0001) and 9 (p = 0.0007) were correlated with carrier isolates and serovar 4 was associated with isolates from animals with respiratory disease (p = 0.015). In systemic isolates, serovar 13 was most frequently detected (18.9%). Various other serovars were isolated from all sites and the ratio of serovar 5 to serovar 12 was approximately 1:2. These two serovars together represented 14.3% of the isolates; only serovar 4 was isolated more frequently (24.7%). The pathotyping method based on the leader sequence (LS = ESPR of vta) was easy to perform and corresponded well to the clinical background information. Of the carrier isolates 72% were identified as non-virulent while 91% of the systemic isolates were classified as virulent (p &amp;lt; 0.0001). Results of the pathotyping PCR based on 10 different marker genes overall were in good agreement with clinical metadata as well as with results of the LS-PCR. However, the pathotyping PCR was more complicated to perform and analyze. In conclusion, a combination of the serotyping multiplex-PCR and the LS-PCR could improve identification of clinically relevant G. parasuis isolates, especially from respiratory samples